A Protein Suspension-Trapping Sample Preparation for Tear Proteomics by Liquid Chromatography-Tandem Mass Spectrometry.

Tear fluid is one of the easily accessible biofluids that can be collected non-invasively. Tear proteomics has the potential to discover biomarkers for several ocular diseases and conditions. The suspension trapping column has been reported to be an efficient and user-friendly sample preparation workflow for the broad application of downstream proteomic analysis. Yet, this strategy has not been well-studied in the analysis of human tear proteome. The present protocol describes an integrated workflow from clinical human tear samples to purified peptides for non-invasive tear protein biomarker research using mass spectrometry, which provides insights into disease biomarkers and monitoring when combined with bioinformatics analysis. A protein suspension trapping sample preparation was applied and demonstrated the discovery of tear proteome with fast, reproducible, and user-friendly procedures, as a universal, optimized sample preparation for human tear fluid analysis. In particular, the suspension trapping procedure outperformed in-solution sample preparation in terms of peptide recovery, protein identification, and shorter sample preparation time.

Data on assessment of safety and tear proteome change in response to orthokeratology lens - Insight from integrating clinical data and next generation proteomics.

Breath-O™ Correct Ortho-K lenses are newly designed ortho-K lenses which are made from a silicon and fluoride containing methacrylate compound. This compound is said to be more flexible, durable and less likely to break compared to traditional Ortho-K lenses. The special design of this Ortho-K lens can reshape the corneal profile to induce temporary myopic reduction while producing beneficial peripheral hyperopic defocus for myopia control. To evaluate the safety and ocular surface responses of overnight Ortho-K wear over 1 and 3 months using this new type of material, we evaluated the clinical parameters (corneal integrity, corneal biomechanics, corneal endothelial health, non-invasive keratographical break-up time) and profiled the change of global tear proteome on healthy young subjects using next generation proteomics (SWATH-MS). The acquired mass spectrometric data were processed and analyzed using a cloud based Oneomics™ bioinformatic platform. All raw data generated from Information-dependent acquisition (IDA) and SWATH acquisitions were accepted and published in the Peptide Atlas public repository for general release (http://www.peptideatlas.org/PASS/PASS01367).